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      Effects of a Chimeric Lysin against Planktonic and Sessile Enterococcus faecalis Hint at Potential Application in Endodontic Therapy

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          Abstract

          Enterococcus faecalis is a commensal opportunistic pathogen found in the intestine, mouth, and vaginal tract of humans. As an invasive pathogen in the oral cavity, E. faecalis is one of the leading causes of periapical endodontic lesions. However, due to the strong biofilm-forming capacity and tolerance of E. faecalis to conventional antibiotics and treatments, limited therapeutic options are available. In the present study, we investigated the activity of ClyR, a chimeric lysin with extended streptococcal lytic spectrum, against planktonic and sessile E. faecalis cells in vitro and in an ex vivo dental model. Our results showed that ClyR has robust and rapid lytic activity against multiple E. faecalis strains, killing >90% planktonic cells within 1 min at a concentration of 50 μg/mL. The biochemical experiments combined with microscopy analysis revealed that ClyR degrades E. faecalis biofilm with high efficacy in a dose-dependent manner, reducing the survival rate to <40% within biofilms after treatment with 50 μg/mL ClyR for 1 h. In the ex vivo dental model, ClyR showed a significant biofilm removal efficacy, killing >90% viable bacteria within biofilms at a low dose of 50 μg/mL, which is much better than ampicillin and similar to calcium hydroxide, the extensively used routine intracanal medicament in the treatment of endodontics and dental traumatology. The robust activity of ClyR against both planktonic and sessile E. faecalis suggests the potential of ClyR in treating endodontic infections caused by E. faecalis.

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          Mechanisms of antimicrobial activity of calcium hydroxide: a critical review.

          Literature review The primary function of calcium hydroxide as a routine intracanal medicament is to provide antimicrobial activity. However, the mechanisms of antimicrobial activity of calcium hydroxide are not well known. Physicochemical properties of this substance may limit its effectiveness in disinfecting the entire root canal system. In addition, calcium hydroxide is not effective against all bacterial species found in root canal infections. Association with other medicaments may enhance the efficacy of the intracanal medication in eliminating residual bacteria in the root canal system.
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            Enterococcus faecalis--a mechanism for its role in endodontic failure.

            Nick Love (2001)
            The aim of this study was to identify a possible mechanism that would explain how E. faecalis could survive and grow within dentinal tubules and reinfect an obturated root canal. Cells of Streptococcus gordonii DL1, Streptococcus mutans NG8, or E. faecalis JH2-2 were grown in brain heart infusion broth containing various amounts of human serum for 56 days. The ability of the three species to invade dentine and bind to immobilized type I collagen in the presence of human serum was assessed by dentine invasion and microtitre well experiments. All three species remained viable over the period of the experiment when grown in human serum. Cells of all three bacteria were able to invade dentine and bind to immobilized collagen. Both of these properties were inhibited by the presence of collagen in the cell solution. Human serum inhibited dentine invasion and collagen adhesion by S. gordonii DL1 and S. mutans NG8, whilst dentine invasion by E. faecalis JH2-2 was reduced in the presence of serum, but not inhibited, and binding to collagen was enhanced. It is postulated that a virulence factor of E. faecalis in failed endodontically treated teeth may be related to the ability of E. faecalis cells to maintain the capability to invade dentinal tubules and adhere to collagen in the presence of human serum.
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              Properties and applications of calcium hydroxide in endodontics and dental traumatology.

              Calcium hydroxide has been included within several materials and antimicrobial formulations that are used in a number of treatment modalities in endodontics. These include, inter-appointment intracanal medicaments, pulp-capping agents and root canal sealers. Calcium hydroxide formulations are also used during treatment of root perforations, root fractures and root resorption and have a role in dental traumatology, for example, following tooth avulsion and luxation injuries. The purpose of this paper is to review the properties and clinical applications of calcium hydroxide in endodontics and dental traumatology including its antibacterial activity, antifungal activity, effect on bacterial biofilms, the synergism between calcium hydroxide and other agents, its effects on the properties of dentine, the diffusion of hydroxyl ions through dentine and its toxicity. Pure calcium hydroxide paste has a high pH (approximately 12.5-12.8) and is classified chemically as a strong base. Its main actions are achieved through the ionic dissociation of Ca(2+) and OH(-) ions and their effect on vital tissues, the induction of hard-tissue deposition and the antibacterial properties. The lethal effects of calcium hydroxide on bacterial cells are probably due to protein denaturation and damage to DNA and cytoplasmic membranes. It has a wide range of antimicrobial activity against common endodontic pathogens but is less effective against Enterococcus faecalis and Candida albicans. Calcium hydroxide is also an effective anti-endotoxin agent. However, its effect on microbial biofilms is controversial. © 2011 International Endodontic Journal.
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                Author and article information

                Journal
                Viruses
                Viruses
                viruses
                Viruses
                MDPI
                1999-4915
                29 May 2018
                June 2018
                : 10
                : 6
                : 290
                Affiliations
                [1 ]The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine, Ministry of Education, School of Stomatology, Wuhan University, Wuhan 430079, China; 2016203040006@ 123456whu.edu.cn
                [2 ]Key Laboratory of Special Pathogens and Biosafety, Center for Emerging Infectious Diseases, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China; wangsj@ 123456wh.iov.cn
                [3 ]State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China; hejin@ 123456mail.hzau.edu.cn
                Author notes
                [* ]Correspondence: yangh@ 123456wh.iov.cn (H.Y.); 1004809372@ 123456whu.edu.cn (Y.L.); hpwei@ 123456wh.iov.cn (H.W.); Tel.: +86-27-5186-1078 (H.Y.); +86-27-8764-7443 (Y.L.); +86-27-5186-1076 (H.W.); Fax: +86-27-8719-9492 (H.Y.); +86-27-8764-7443 (Y.L.); +86-27-8719-9492 (H.W.)
                [†]

                These authors contributed equally to this work.

                Author information
                https://orcid.org/0000-0001-6750-1465
                Article
                viruses-10-00290
                10.3390/v10060290
                6024690
                29844267
                66b0d778-f71e-4800-aa85-acdb7a3e5890
                © 2018 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 29 April 2018
                : 23 May 2018
                Categories
                Article

                Microbiology & Virology
                bacteriophage lysin,enterococcus faecalis,bacterial biofilm,endodontic infection,calcium hydroxide

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